Following causes can be excluded:

- too little sample, enzymkojugate, substrate and stop solution

- too little exposure (reaction) time

- wrong handling with the pipette

- too less warm up time after removal from the cooling area

- false photometer settings

- ELISA-Kit used after the expiration date

- any step that is connected with disregarding manufacturer's specifications

The ELISA-Kit was carried out manually.

Enzymconjugate, substrate and stop solution was applied with an electrical pipette, whose battery isn't working well what leads sometimes to problems. So I suppose, that the pipette could be one source of error.

Could you please help me, to find out possible other causes?

I would be gratful for any advice or help, thank you!

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